Compositions and methods for controlling arthropod parasite and pest infestations
Inventors
Inberg, Alex • Kapoor, Mahak • Evans, Jay
Assignees
United States, AGRICULUTURE, Secretary of • US Department of Agriculture USDA • Greenlight Biosciences Inc
Publication Number
US-9540642-B2
Publication Date
2017-01-10
Expiration Date
2034-11-04
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Abstract
This application provides and discloses anti-parasitic, anti-pest or insecticidal nucleic acid molecules and their calmodulin target genes for the control of arthropod parasites and pests. This application further provides methods and compositions for the control and treatment of parasites and pests in Apis mellifera (honey bee) hives.
Core Innovation
This invention provides anti-parasitic, anti-pest, or insecticidal nucleic acid molecules targeting calmodulin genes of arthropod parasites and pests, particularly for controlling Varroa mite infestations in honey bee (Apis mellifera) hives. The compositions include nucleic acid molecules, such as dsRNA or siRNA, that are essentially identical or complementary to regions of calmodulin gene sequences from the target parasite, with excipients suitable for delivering these nucleic acids to bees or mites.
The problem addressed relates to the significant threat that arthropod pests and parasites, notably Varroa destructor mites, pose to arthropod culture and specifically to honey bee colonies critical for agriculture. Existing Varroa control methods, including chemical miticides and management practices, lose efficacy, bear toxicity, and risk contamination of honey products. There is a lack of safe, effective, and durable solutions to prevent or treat Varroa infestation and consequently Colony Collapse Disorder (CCD).
The invention overcomes these challenges by providing selective insecticide compositions that suppress calmodulin gene expression in Varroa mites via RNA interference, resulting in mite mortality or reduced reproduction. The compositions can be administered via bee feed or topical application, allowing uptake and systemic distribution of the nucleic acid molecules to the parasites through the host. The compositions are designed to be selective to the parasite calmodulin genes, minimizing off-target effects on the host bee or other non-target organisms.
Claims Coverage
The patent includes 15 claims encompassing compositions, nucleic acid constructs, and methods of use focused on nucleic acid molecules targeting Varroa destructor calmodulin genes.
Selective nucleic acid composition for bees and mites
A composition that is bee-ingestible, bee-absorbable, mite-ingestible, or mite-absorbable comprising an excipient and a nucleic acid molecule having at least 96% identity or complementarity to at least 21 contiguous nucleotides of SEQ ID NO: 3.
Use of double-stranded RNA in compositions
The nucleic acid molecule in the composition can be dsRNA, particularly siRNA, targeting specific regions of the calmodulin gene.
Composition excipients
The excipient in the composition is selected from proteins, pollen, carbohydrates, polymers, liquid solvents, sugar syrups, sugar solids, or semi-solid feed to facilitate ingestion or absorption.
Nucleic acid constructs for dsRNA production
A nucleic acid construct comprising a nucleic acid encoding a sequence at least 95% identical or complementary to SEQ ID NO: 3 operably linked to a promoter functional in a host cell to produce dsRNA, optionally including regulatory elements such as introns or enhancers, and applicable in bacterial or yeast cells.
Methods of administration to honeybees
Methods of administering an effective amount of the composition to honeybees, resulting in the presence of the nucleic acid in bee tissue, where the calmodulin gene sequence targeted is from Varroa destructor, reducing susceptibility, parasitation, or parasite load in the colony or individual bees.
Compositions with multiple nucleic acid molecules
The composition may further include one or more nucleic acid molecules complementary or identical to different regions of a calmodulin gene to enhance anti-parasitic action.
The claims cover selective nucleic acid compositions comprising dsRNA targeting Varroa destructor calmodulin genes, nucleic acid constructs for dsRNA expression in host cells, methods of administering these compositions to bees, and compositions with multiple nucleic acid molecules targeting distinct calmodulin gene regions for effective control of Varroa mite infestation.
Stated Advantages
Provides a safe, efficacious, and long-lasting solution to control Varroa destructor mites in honey bee colonies.
Selective insecticide compositions minimize toxicity and residue issues in honey and beeswax compared to chemical miticides.
Methods reduce parasitation and parasite load of Varroa destructor, thereby potentially preventing or treating Colony Collapse Disorder in bee colonies.
Compositions enable RNA interference targeting parasite-specific calmodulin genes, resulting in effective mite mortality and suppression of reproduction.
Excipients facilitate delivery and uptake of nucleic acids by bees and mites, ensuring practical applicability in beekeeping.
Documented Applications
Controlling and treating Varroa destructor mite infestations in honey bee colonies.
Reducing susceptibility of individual honeybees and bee colonies to Varroa destructor parasitation.
Preventing or treating Colony Collapse Disorder through Varroa mite control.
Reducing parasite load in commercial bee hives by providing compositions via bee feed or topical application.
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